基于靶向胆汁酸代谢组学探索实热证胆结石病发病预警信号

Based on our 3-year longitudinal data in a community in Beijing, we indicated that there were some specific changes in serum bile acid (BA) profile in 6-12 months before the patients could be diagnosed with gallstone disease and excessive heat syndrome (EHS) in traditional Chinese medicine (TCM). These changes might be potential predictive markers for the future development of gallstone, thus to provide a possibility for clinicians to try to prevent the formation of gallstone at an earlier time. Thus in this study, we planned to analyzed serum BA profiles from healthy adults and patients with GS diagnosis over 12 months and patients from their Pre-GS period until neodeveloped GS stage by using a targeted metabonomics approach with ultraperformance liquid chromatography triple−quadrupole mass spectrometry. The patients undergo their formation of GS should be diagnosed with no other diseases and medicinal history so as to lower the perturb as far as possible. The three groups of individuals should be matched by age, gender, height, and weight at a 1:1:1 ratio. Then try to detect the specific changes of BA profiles in Pre-GS group compared with other groups, as the prewarning markers for EHS-GS development within 6-12 months. Then the identified markers will be validated in an independent realworld cohort with a very high incidence rate of GS. The study aims at providing a better understanding of GS development, as well as fine indicators to prevent the formation of GS in a proper earlier stage.

在前期基于社区人群的连续3年随访中，通过对新发实热证胆结石病患者的初步观察，发现实热证胆结石病患者早在发病前6-12个月即发生胆汁酸水平的特异性变化，可能为后期的疾病发生提供准确的预警信息，从而使疾病的早期预防性干预成为可能。.本研究基于已建立的血清样本库，拟选择排除其他疾病干扰且未服用相关药物的新发实热证胆结石病的患者，收集其发病前6-12个月、和发病后6个月内的血清样本，根据年龄、性别、身高、体重匹配健康人、胆结石病发病1年以上患者作为对照，应用靶向代谢组学方法检测各组血清胆汁酸代谢谱，探索胆汁酸谱的特异性变化，找到实热证胆结石病患者发病的预警性血清胆汁酸信号；并在另一独立的、基于真实世界的胆结石病高发队列人群中进行验证。从而深化对胆结石病发病机制的理解，并为后期临床早期干预结石形成、恰当选择预防指征和时间节点提供依据。

本研究以新发实热证胆结石病（排除其他疾病干扰且未服用药物治疗）为例，探索该病发生前期的血清胆汁酸谱特异性表征，并进行临床验证，以建立疾病早期预警、防治的新方法。.基于课题组前期建立的临床样本库，筛选新发实热证胆结石病患者20例，根据年龄、性别、BMI匹配健康人和胆结石病发病12个月以上患者各30例作为对照，收集其发病前6-12个月和发病后12个月内的血清样本，应用靶向代谢组学方法检测各组血清胆汁酸代谢谱，分析胆汁酸谱的特异性变化，提取发病的预警信号，并在另一独立的胆结石病高发队列——接受胃切除手术的2型糖尿病患者队列（胆结石发病率21%）中进行验证性分析。研究发现，在胆结石病发生的前6-12个月，体内的胆汁酸水平已经开始发生变化；胆汁酸谱的特异性改变可以有效区别健康人、胆结石前期、新发胆结石患者、和长期胆结石患者各组人群。8种胆汁酸组合的变化可以较好的区分四组，包括HDCA, 6-KetoLCA, 7-KetoDCA, THDCA, HCA, TBA, GCA, TCA。验证性分析发现，TLCA和TBA的组合变化率及其分别变化率、TLCA/GLCA比值对于未来12个月内胆结石病的发生均具有较好的预警价值。TLCA和TBA组合指标在验证队列的AUC曲线下面积为0.815；敏感性和特异性分别为0.842和0.789。TLCA和TBA单独变化AUC分别为0.832和0.770。TLCA/GLCA比值的AUC为0.696。.同时对于胆汁酸在胆结石病患者中的变化研究开展全面、系统性文献研究，定性研究发现TCA、GCDCA、GCA、TCDCA、GDCA、DCA在胆结石病人血清中显著升高；包含16个研究的meta分析显示，胆结石病患者血清TBA升高，而胆汁中TBA降低；血清GCA、血清TCA、胆汁TCDCA升高、胆汁GCDCA降低，对于本研究结果给予文献支持。开展的小分子代谢物/胆汁酸分子相关的疾病-药物药理网络构建与分析方法学探索，为下一步干预性研究奠定基础。.完成学术论文12篇，与本项目直接相关的论文已发表3篇（SCI 收录2 篇，中文1篇）；另2篇已投稿SCI期刊（1篇已修回，1篇审稿过程中）；1篇待投稿。参加国内学术会议交流2次；相关学术培训3次；培养硕士研究生2名。